Novel Structure of the Conserved Gram-Negative Lipopolysaccharide Transport Protein A and Mutagenesis Analysis

Lipopolysaccharide (LPS) transport protein A (LptA) is an essential periplasmic localized transport protein that has been implicated together with MsbA, LptB, and the Imp/RlpB complex in LPS transport from the inner membrane to the outer membrane, thereby contributing to building the cell envelope i...

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Veröffentlicht in:Journal of molecular biology 2008-07, Vol.380 (3), p.476-488
Hauptverfasser: Suits, Michael D.L., Sperandeo, Paola, Dehò, Gianni, Polissi, Alessandra, Jia, Zongchao
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container_issue 3
container_start_page 476
container_title Journal of molecular biology
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creator Suits, Michael D.L.
Sperandeo, Paola
Dehò, Gianni
Polissi, Alessandra
Jia, Zongchao
description Lipopolysaccharide (LPS) transport protein A (LptA) is an essential periplasmic localized transport protein that has been implicated together with MsbA, LptB, and the Imp/RlpB complex in LPS transport from the inner membrane to the outer membrane, thereby contributing to building the cell envelope in Gram-negative bacteria and maintaining its integrity. Here we present the first crystal structures of processed Escherichia coli LptA in two crystal forms, one with two molecules in the asymmetric unit and the other with eight. In both crystal forms, severe anisotropic diffraction was corrected, which facilitated model building and structural refinement. The eight-molecule form of LptA is induced when LPS or Ra-LPS (a rough chemotype of LPS) is included during crystallization. The unique LptA structure represents a novel fold, consisting of 16 consecutive antiparallel β-strands, folded to resemble a slightly twisted β-jellyroll. Each LptA molecule interacts with an adjacent LptA molecule in a head-to-tail fashion to resemble long fibers. Site-directed mutagenesis of conserved residues located within a cluster that delineate the N-terminal β-strands of LptA does not impair the function of the protein, although their overexpression appears more detrimental to LPS transport compared with wild-type LptA. Moreover, altered expression of both wild-type and mutated proteins interfered with normal LPS transport as witnessed by the production of an anomalous form of LPS. Structural analysis suggests that head-to-tail stacking of LptA molecules could be destabilized by the mutation, thereby potentially contributing to impair LPS transport.
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subjects Amino Acid Sequence
anisotropic diffraction
Anisotropy
Biological Transport - physiology
Carrier Proteins - chemistry
Carrier Proteins - metabolism
Crystallography, X-Ray
Escherichia coli
Escherichia coli - genetics
Escherichia coli - metabolism
Escherichia coli - ultrastructure
lipopolysaccharide transport
Lipopolysaccharides - analysis
Lipopolysaccharides - chemistry
Lipopolysaccharides - isolation & purification
Lipopolysaccharides - metabolism
LptA and YhbN
Models, Biological
Models, Chemical
Molecular Sequence Data
Mutagenesis, Site-Directed
periplasm
Protein Folding
Protein Structure, Secondary
Recombinant Proteins - chemistry
Recombinant Proteins - isolation & purification
Recombinant Proteins - metabolism
Sequence Homology, Amino Acid
Spectrum Analysis, Raman
X-ray crystal structure
X-Ray Diffraction
title Novel Structure of the Conserved Gram-Negative Lipopolysaccharide Transport Protein A and Mutagenesis Analysis
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